Microfluidic continuous flow digital loop-mediated isothermal amplification (LAMP).
Microfluidic continuous flow digital loop-mediated isothermal amplification (LAMP).
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DOI：
10.1039/c4lc01158a
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发表时间：
2015-02-07
期刊：
影响因子：
6.1
通讯作者：
Wang TH
中科院分区：
文献类型：
--
作者：
Rane TD;Chen L;Zec HC;Wang TH
数字核酸检测正在迅速成为广泛生物医学研究中核酸分子超灵敏定量检测的流行技术。数字聚合酶链式反应 (PCR) 仍然是进行数字核酸检测的最流行的方法。然而，由于热循环的需要，数字PCR很难在单个微流控设备上以简化的方式实施，导致复杂的支离破碎的工作流程和多个独立的设备和仪器。环介导等温扩增 (LAMP) 是 PCR 的绝佳等温替代方案，通过对核酸靶标使用多个引物组，具有比 PCR 更好的特异性。在这里，我们报告了一种微流控液滴装置，该装置实现了数字核酸检测所需的所有步骤，包括数字 LAMP 的液滴生成、孵育和在线检测。与基于微室或液滴阵列的数字化验相比，该设备的连续流动操作消除了设备占地面积对总反应数量的限制，以及仪器之间长时间孵育和材料传输对分析吞吐量的限制。
Digital nucleic acid detection is rapidly becoming a popular technique for ultra-sensitive and quantitative detection of nucleic acid molecules in a wide range of biomedical studies. Digital polymerase chain reaction (PCR) remains the most popular way of conducting digital nucleic acid detection. However, due to the need for thermocycling, digital PCR is difficult to implement in a streamlined manner on a single microfluidic device, leading to complex fragmented workflows and multiple separate devices and instruments. Loop-mediated isothermal amplification (LAMP) is an excellent isothermal alternative to PCR with potentially better specificity than PCR through the use of multiple primer sets for a nucleic acid target. Here we report a microfluidic droplet device implementing all the steps required for digital nucleic acid detection including droplet generation, incubation and in-line detection for digital LAMP. As compared to microchamber or droplet array-based digital assays, continuous flow operation of this device eliminates the constraints on the number of total reactions by the footprint of the device and the analysis throughput by the time for lengthy incubation and transfers of materials between instruments.